Arrays of discrete cell culture microenvironments, methods of making such arrays and uses thereof

Abstract

A method of making an array having discrete volumes of cell culture micro-environments possessing different properties influencing the behavior of encapsulated cells, in particular proliferation, colony-formation, differentiation, migration, or combinations thereof. The method comprises a) providing different hydrogel precursor molecules, where the precursor molecules are linear or branched hydrophilic polymers for building up cell culture micro-environments; b) combining and dispensing different combinations of the hydrogel precursor molecules, according to step a), onto discrete volumes of a multi-well plate; c) adding one or more biologically active molecules to the discrete volumes and either attaching the molecules to one of the hydrogel precursor molecules present or a hydrogel, formed in step e), or allowing them to diffuse freely; d) adding cells onto / into the discrete volumes of the multi-well plate; and e) crosslinking the hydrogel precursor molecules based on an enzymatically catalyzed reaction or a Michael addition reaction to form a hydrogel matrix.

Description

[0001]The present invention pertains to arrays of discrete volumes of cell culture microenvironments possessing different properties, i.e. different mechanical and / or biochemical properties (i.e. cell adhesion, signalling, degradability) influencing cell phenotype and fate, i.e. proliferation / self-renewal, colony formation, differentiation and / or migration, methods of making such arrays, a kit-of-parts for making such arrays and a combinatorial method of testing the influence of hydrogel formulations on cell growth behaviour, in particular of difficult-to-culture cell types such as stem cells.BACKGROUND OF THE INVENTION[0002]Two-dimensional cell culture systems as well as naturally derived three-dimensional cell culture systems as models to elucidate complex cell behaviour, notably in the field of cancer or stem cell research, have been known for many years.[0003]Two-dimensional cell culture systems have the drawback of not allowing for gene expression patterns and cellular phenotyp...

Images